降解植物甾醇产雄甾1,4-二烯-3,17-二酮工程菌株的构建及转化培养基优化  

Construction of Engineering Bacteria for Degrading Phytosterol to Androst-1,4-diene-3,17-dione and the Optimization of Transformation Medium

作　　者：刘相岑 郝晓蔚[3] 张瑞婕[2] 冯雯雯 朱晨光[1] 张保国[2] 史吉平[2] LIU Xiang-cen;HAO Xiao-wei;ZHANG Rui-jie;FENG Wen-wen;ZHU Chen-guang;ZHANG Bao-guo;SHI Ji-pingz(College of Life Sciences,Shanghai University,Shanghai 200444,China;Shanghai Advanced Research Institute,Chinese Academy of Sciences,Shanghai 201210,China;Hebei University of Science and Technology,Shijiazhuang 05001g,China)

机构地区：[1]上海大学生命科学学院,上海200444 [2]中国科学院上海高等研究院,上海201210 [3]河北科技大学,河北石家庄050018

出　　处：《食品工业科技》2018年第18期110-116,共7页Science and Technology of Food Industry

基　　金：国家自然科学基金(41106125)

摘　　要：雄甾-1,4-二烯-3,17-二酮(ADD)作为甾体激素药物不可替代的中间体,是多种激素类药物的原材料。在分枝杆菌(Mycobacterium sp.ZFZ)体内过表达3-甾酮-Δ1脱氢酶(Kst D),获得一株ADD生产菌,随后研究了不同培养基组成对该菌株生产ADD的影响。在单因素的基础上,以ADD产量为衡量指标,采用响应面法优化了转化培养基的组成,并建立各影响因素变化的二次回归方程。结果表明,最适宜的转化培养条件为玉米浆23 g/L、葡萄糖9 g/L、硝酸钠2.8 g/L、磷酸氢二钾2 g/L。此条件下底物甾醇浓度为10 g/L时,ADD的产量可达(4.12±0.18)g/L,ADD产量比出发菌株提高了近18倍。Androsten-1,4-diene-3,17-dione（ADD）was an irreplaceable intermediate and a raw material for steroid hormone drugs.A strain of ADD producing bacteria was obtained by overexpressing 3-sterosterone-Δ-1-dehydrogenase（Kst D）in Mycobacterium sp.ZFZ.Subsequently,the effects of different media compositions on the production of ADD by this strain were studied.On the basis of single factor experiments,ADD production as index response surface analysis was applied to optimize the fermentation medium composition.The quadratic regression analysis was applied to get the optimal level of main factors.The results showed that the most suitable transformation culture conditions were corn steep liquor 23 g/L,glucose 9 g/L,Na NO32.8 g/L and K2HPO42 g/L.The yield of ADD reached（4.12±0.18）g/L when the concentration of substrate sterol was10 g/L in the optimal condition.And the yield of ADD increased nearly 18 times than Mycobacterium sp.ZFZ.

关 键 词：植物甾醇 生物转化 3-甾酮-Δ1脱氢酶 雄甾-1 4-二烯-3 17-二酮 

分 类 号：TS201.3[轻工技术与工程—食品科学]